Plants and seeds of common wheat cultivar WB-112

ABSTRACT

A wheat cultivar, designated WB-112, is disclosed. The invention relates to the seeds, plants, and hybrids of wheat cultivar WB-112, and to methods for producing a wheat plant produced by crossing plants from wheat cultivar WB-112 with themselves or with plants from another wheat variety. The invention also relates to methods for producing a wheat plant containing in its genetic material one or more transgenes and to the transgenic wheat plants and plant parts produced by those methods. The invention also relates to wheat varieties or breeding varieties and plant parts derived from wheat cultivar WB-112, to methods for producing other wheat varieties, lines or plant parts derived from wheat cultivar WB-112, and to the wheat plants, varieties, and their parts derived from the use of those methods. The invention further relates to hybrid wheat seeds and plants produced by crossing wheat cultivar WB-112 with another wheat cultivar.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates generally to a new and distinctive wheatline, designated WB-112, representative sample of seed of which wasdeposited under ATCC Accession No. PTA-11955, as well as to seed,plants, cultivars and hybrids related thereto. The invention alsorelates to methods for producing wheat seed and plants from WB-112.

SUMMARY OF THE INVENTION

In an embodiment, the invention is directed to a seed, wheat plant orpart thereof, or tissue culture of wheat cultivar WB-112. The inventionis also directed, in an embodiment, to a method for producing a wheatseed comprising crossing two wheat plants and harvesting the resultantwheat seed, wherein at least one of the two wheat plants is from theWB-112 line. The invention is also directed, in various embodiments, tomethods of producing plants and plants that are herbicide tolerant, pestor insect resistant, disease resistant, or have modified fatty acid orcarbohydrate metabolisms.

In yet another embodiment, the invention is directed to a method ofintroducing a desired trait into wheat cultivar WB-112, wherein themethod comprises:

-   -   (a) crossing a WB-112 plant with a plant of another wheat        cultivar that comprises a desired trait to produce progeny        plants wherein the desired trait is selected from the group        consisting of male sterility, herbicide tolerance, herbicide        resistance, insect resistance, modified fatty acid metabolism,        modified carbohydrate metabolism, modified protein metabolism,        modified phytic acid metabolism, modified waxy starch content,        modified protein content, increased tolerance to water stress        and resistance to bacterial disease, fungal disease or viral        disease;    -   (b) selecting one or more progeny plants that have the desired        trait to produce selected progeny plants;    -   (c) crossing the selected progeny plants with the WB-112 plant        to produce backcross progeny plants;    -   (d) selecting for backcross progeny plants that have the desired        trait and essentially all of the physiological and morphological        characteristics of wheat cultivar WB-112 listed in Table 1; and    -   (e) repeating the crossing the selected progeny step and        selecting for backcross progeny step two or more times in        succession to produce selected third or higher backcross progeny        plants that comprise essentially all of the desired trait and        all of the physiological and morphological characteristics of        wheat cultivar WB-112 listed in Table 1.

DEFINITIONS

The following definitions apply to the terms used herein:

Allele. Allele is any of one or more alternative forms of a gene, all ofwhich relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Awn. Awn is intended to mean the elongated needle-like appendages on theflower- and seed-bearing head at the top of the cereal grain plant(e.g., wheat, common wheat, rye). Awns are attached to the lemmas.Lemmas enclose the stamen and the stigma as part of the florets. Floretsare grouped in spikelets, which in turn together comprise the head.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents. For example, a firstgeneration hybrid F₁ may be crossed with one of the parental genotypesof the F₁ hybrid.

Cell. As used herein, the term cell includes a plant cell, whetherisolated, in tissue culture, or incorporated in a plant or plant part.

Disease Resistance. As used herein, the term disease resistance ordisease resistant is defined as the ability of plants to restrict theactivities of a specified disease, such as a fungus, virus, orbacterium.

Disease Tolerance. As used herein, the term disease tolerance or diseasetolerant is defined as the ability of plants to endure a specifieddisease (such as a fungus, virus or bacterium) or an adverseenvironmental condition and still perform and produce in spite of thisdisorder.

Embryo. The embryo is the small plant contained within a mature seed.

Essentially all of the physiological and morphological characteristics.This phrase refers to a plant having essentially all of thephysiological and morphological characteristics of the referenced plantor variety, as determined at a 5% significance level for quantitativedata.

Gene. As used herein, gene refers to a segment of nucleic acid. A genecan be introduced into a genome of a species, whether from a differentspecies or from the same species, using transformation or variousbreeding methods.

Gene Converted (Conversion). Gene conversion or a gene converted plantrefers to plants that are developed by backcrossing, geneticengineering, or mutation, wherein essentially all of the desiredmorphological and physiological characteristics of a variety arerecovered, in addition to the one or more traits transferred into thevariety via the backcrossing technique, genetic engineering, ormutation.

Gene Silencing. Gene silencing means the interruption or suppression ofthe expression of a gene at the level of transcription or translation.

Genotype. Genotype refers to the genetic constitution of a cell ororganism.

Head. As used herein, the term head refers to a group of spikelets atthe top of one plant stem. The term spike also refers to the head of aplant located at the top of one plant stem.

Glume Blotch. Glume Blotch is a disease of wheat characterized by small,irregular gray to brown spots or blotches on the glumes, althoughinfections may also occur at the nodes. The disease is caused by thefungus Stagonosporum nodorum (may also be referred to as Septorianodorum). Resistance to this disease is scored on scales that reflectthe observed extent of the disease on the leaves of the plant. Ratingscales may differ but in general a low number indicates resistance andhigher number suggests different levels of susceptibility.

Herbicide Resistance. As used herein, the term herbicide resistance orherbicide resistant is defined as the ability of plants to survive andreproduce following exposure to a dose of herbicide that would normallybe lethal to the plant.

Herbicide Tolerance. As used herein, the term herbicide tolerance orherbicide tolerant is defined as the ability of plants to survive andreproduce after herbicide treatment.

Insect Resistance. As used herein, the term disease resistance ordisease resistant is defined as the ability of plants to restrict theactivities of a specified insect or pest.

Insect Tolerance. As used herein, the term disease tolerance or diseasetolerant is defined as the ability of plants to endure a specifiedinsect or pest and still perform and produce in spite of this disorder.

Kernel Weight. As used herein, the term kernel weight refers to theweight of individual kernels (also called seeds), often reported as theweight of one thousand kernels or “1000 Kernel Weight”.

Leaf Rust. Leaf Rust is a disease of wheat characterized by pustulesthat are circular or slightly elliptical, that usually do not coalesce,and contain masses of orange to orange-brown spores. The disease iscaused by the fungus Puccinia recondita f. sp. tritici. Infection sitesprimarily are found on the upper surfaces of leaves and leaf sheaths,and occasionally on the neck and awns. Resistance to this disease isscored on scales that reflect the observed extent of the disease on theleaves of the plant. Rating scales may differ but in general a lownumber indicates resistance and higher number suggests different levelsof susceptibility.

Linkage. Linkage refers to a phenomenon wherein alleles on the samechromosome tend to segregate together more often than expected if theirtransmission was independent.

Locus. A locus is a position on a genomic sequence that is usually foundby a point of reference, for example, the position of a DNA sequencethat is a gene, or part of a gene or intergenic region. A locus confersone or more traits such as, for example, male sterility, herbicidetolerance or resistance, insect resistance or tolerance, diseaseresistance or tolerance, modified fatty acid metabolism, modified phyticacid metabolism, modified carbohydrate metabolism or modified proteinmetabolism. The trait may be, for example, conferred by a naturallyoccurring gene introduced into the genome of the variety bybackcrossing, a natural or induced mutation, or a transgene introducedthrough genetic transformation techniques. A locus may comprise one ormore alleles integrated at a single chromosomal location.

Maturity. As used herein, the term maturity refers to the stage of plantgrowth at which the development of the kernels is complete.

Pedigree Distance. Pedigree distance is the relationship amonggenerations based on their ancestral links as evidenced in pedigrees. Itmay be measured by the distance of the pedigree from a given startingpoint in the ancestry.

Percent Identity. Percent identity, as used herein, refers to thecomparison of the homozygous alleles of two wheat varieties. Percentidentity is determined by comparing a statistically significant numberof the homozygous alleles of two developed varieties. For example, apercent identity of 90% between wheat variety 1 and wheat variety 2means that the two varieties have the same allele at 90% of their loci.

Percent Similarity. Percent similarity as used herein refers to thecomparison of the homozygous alleles of a wheat variety such as WB-112with another plant, and if the homozygous allele of WB-112 matches atleast one of the alleles from the other plant then they are scored assimilar. Percent similarity is determined by comparing a statisticallysignificant number of loci and recording the number of loci with similaralleles as a percentage. A percent similarity of 90% between WB-112 andanother plant means that WB-112 matches at least one of the alleles ofthe other plant at 90% of the loci.

Plant. As used herein, the term plant includes reference to an immatureor mature whole plant, including a plant from which seed, grain, oranthers have been removed. A seed or embryo that will produce the plantis also considered to be a plant.

Plant Height (Hgt). As used herein, the term plant height is defined asthe average height in inches or centimeters of a group of plants, asmeasured from the ground level to the tip of the head, excluding awns.

Plant Parts. As used herein, the term plant parts (or reference to “awheat plant, or a part thereof”) includes but is not limited toprotoplasts, callus, leaves, stems, roots, root tips, anthers, pistils,seed, grain, pericarp, embryo, pollen, ovules, cotyledon, hypocotyl,spike, floret, awn, lemma, shoot, tissue, petiole, cells, andmeristematic cells.

Powdery Mildew. Powdery Mildew is a disease of wheat characterized bywhite to pale gray, fuzzy or powdery colonies of mycelia, and conidia onthe upper surfaces of leaves and leaf sheaths (especially on lowerleaves), and sometimes on the spikes. The disease is caused by thefungus Erysiphe graminis f. sp. tritici. Older fungal tissue isyellowish gray. This superficial fungal material can be rubbed offeasily with the fingers. Host tissue beneath the fungal material becomeschlorotic or necrotic and, with severe infections, the leaves may die.Eventually, black spherical fruiting structures may develop in themycelia, and can be seen without magnification. Resistance to thisdisease is scored on scales that reflect the observed extent of thedisease on the leaves of the plant. Rating scales may differ but ingeneral a low number indicates resistance and higher number suggestsdifferent levels of susceptibility.

Progeny. As used herein, progeny includes an F₁ wheat plant producedfrom the cross of two wheat plants where at least one plant includeswheat cultivar WB-112. Progeny further includes but is not limited tosubsequent F₂, F₃, F₄, F₅, F₆, F₇, F₈, F₉ and F₁₀ generational crosseswith the recurrent parental line.

Quantitative Trait Loci (QTL). Quantitative trait loci refer to geneticloci that control to some degree numerically representable traits thatare usually continuously distributed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Rhizoctonia Root Rot. Rhizoctonia Root Rot is a disease of wheatcharacterized by sharp eyespot lesions that develop on basal leafsheaths. The disease is caused by the fungus Rhizoctonia solani. Thelesion margins are dark brown with pale, straw-colored centers and themycelia often present in the centers of lesions are easily removed byrubbing. Roots can also be affected, usually becoming brown in color andreduced in number. The disease can cause stunting and a reduction in thenumber of tillers. Resistance to this disease is scored on scales thatreflect the observed extent of the disease on the leaf sheaths of theplant and on reduced vigor of the plant. Rating scales may differ but ingeneral a low number indicates resistance and higher number suggestsdifferent levels of susceptibility.

Scab or Head Blight. Scab or Head Blight a disease of wheatcharacterized by florets (especially the outer glumes) that becomeslightly darkened and oily in appearance. The disease is caused by thefungus Fusarium which has numerous species. Spores are produced that cangive the spike and shriveled, infected kernels a bright pinkish color.Spores can produce a toxin, deoxynivalenol which can be measured with achemical test. Resistance to this disease can be measured in three ways:the extent of the disease on the spikes of the plant, the percentkernels which are visibly shriveled and the amount of deoxynivalenol inthe kernels. Rating scales may differ but in general a low numberindicates resistance and higher number suggests different levels ofsusceptibility.

SDS Sedimentation. SDS sedimentation (sodium dodecyl sedimentation) testvalues are a measure of the end-use mixing and handling properties ofbread dough and their relation to bread-making quality as a result ofthe dough's gluten quality. Higher SDS sedimentation levels reflecthigher gluten quality.

Septoria Leaf Blotch or Speckled Leaf Blotch. Speckled leaf blotch is adisease of wheat, common wheat and durum wheat characterized byirregularly shaped blotches that are at first yellow and then turnreddish brown with grayish brown dry centers, caused by the rust fungusSeptoria tritici. Resistance to this disease is scored on scales thatreflect the observed extent of the disease on the leaves of the plant.Rating scales may differ but in general a low number indicatesresistance and higher number suggests different levels ofsusceptibility.

Soil Born Mosaic Virus. Soil born mosaic virus is a disease of wheatcharacterized by mild green to yellow mosaic, yellow-green mottling,dashes, and parallel streaks, most clearly visible on the youngest leaf.Reddish streaking and necrosis at leaf tips sometimes occurs. Stuntingcan be moderate to severe, depending on the variety. The disease iscaused by a virus which is transmitted by a soilborne fungus-likeorganism, Polymyxa graminis, which makes swimming spores that infect theroots of wheat. Resistance to this disease is scored on scales thatreflect the observed extent of the disease on the young plants. Ratingscales may differ but in general a low number indicates resistance andhigher number suggests different levels of susceptibility.

Stem Rust. Stem Rust is a disease of wheat characterized by pustulescontaining masses of spores that are dark reddish brown, and may occuron both sides of the leaves, on the stems, and on the spikes. Thedisease is caused by the fungus Puccinia graminis f. sp. Tritici.Resistance to this disease is scored on scales that reflect the observedextent of the disease on the leaves of the plant. Rating scales maydiffer but in general a low number indicates resistance and highernumber suggests different levels of susceptibility.

Stripe Rust. Stripe rust is a disease of wheat, common wheat, durumwheat, and barley characterized by elongated rows of yellow spores onthe affected parts, caused by a rust fungus, Puccinia striiformis.Resistance to this disease is scored on scales that reflect the observedextent of the disease on the leaves of the plant. Rating scales maydiffer but in general a low number indicates resistance and highernumber suggests different levels of susceptibility.

Test Weight (TWT). As used herein, the term test weight is a measure ofdensity that refers to the weight in pounds of the amount of kernelscontained in a bushel unit of volume.

Tissue Culture. As used herein, the term tissue culture indicates acomposition comprising isolated cells of the same or a different type ora collection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollen, ovules, pericarp, flowers,florets, heads, spikelets, seeds, leaves, stems, roots, root tips,anthers, pistils, awns, stems, and the like.

DETAILED DESCRIPTION OF THE INVENTION

Reference now will be made in detail to the embodiments of theinvention, one or more examples of which are set forth below. Eachexample is provided by way of explanation of the invention, not alimitation of the invention. In fact, it will be apparent to thoseskilled in the art that various modifications and variations can be madein the present invention without departing from the scope or spirit ofthe invention. For instance, features illustrated or described as partof one embodiment, can be used on another embodiment to yield a stillfurther embodiment.

Thus, it is intended that the present invention covers suchmodifications and variations as come within the scope of the appendedclaims and their equivalents. Other objects, features and aspects of thepresent invention are disclosed in or are obvious from the followingdetailed description. It is to be understood by one of ordinary skill inthe art that the present discussion is a description of exemplaryembodiments only, and is not intended as limiting the broader aspects ofthe present invention.

Six main wheat market classes exist, five of which belong to the speciesTriticum aestivum L.: common wheat, hard red winter, hard red spring,soft red winter, and white. The sixth class of wheat is durum (Triticumturgidum L.). Common wheats are used in a variety of food products suchas bread, cookies, cakes, crackers, and noodles. In general, the hardwheat classes are milled into flour used for breads and the soft wheatclasses are milled into flour used for pastries and crackers. Wheatstarch is used in the food and paper industries, as laundry starches,and in other products. Products produced from wheat may include grain,flour, baked goods, cereals, pasta, beverages, livestock feed, biofuel,straw, construction materials, and starches.

In an embodiment, the invention is directed to wheat cultivar WB-112,its seeds, plants, and hybrids. Wheat cultivar WB-112 is a soft red,winter type common wheat bred for fall and winter planting in the softred winter wheat growing regions of the United States. The primary usageof wheat cultivar WB-112 will be for production of grain, but it canalso be used for production of silage harvested in the soft dough stage.

A. Origin and Breeding History

Wheat cultivar WB-112 is a common wheat variety developed from a F1selection made from the initial cross of “P2548//STINE 481/OH413/W753”.“P2548” is a proprietary common wheat variety of Pioneer Hi-BredInternational Inc., “Stine 481” is a proprietary common wheat of MidWestOilseeds Inc., “OH413” is a proprietary common wheat of The Ohio StateUniversity and “W753” is a proprietary common wheat of MidWest OilseedsInc. The F1 selection was self pollinated and a single seed descent andmodified bulk system of breeding were used to develop and select WB-112which is an F4 level selection. Some of the criteria used for selectionin various generations include yield, test weight, milling score, strawstrength, maturity, disease resistance, and plant height.

Wheat cultivar WB-112 has shown uniformity and stability in appearanceand performance across several generations (F7-F10) and acrossenvironments where it has been tested. The variety has been increasedwith continued observation for uniformity of plant type as described inthe following variety description information.

B. Phenotypic Description

In accordance with another aspect of the invention, there is provided awheat plant having the physiological and morphological characteristicsof WB-112 as presented in Table 1.

TABLE 1 Physiological and Morphological Characteristics of WB-112VALUES/RATINGS CHARACTERISTIC WB-112 Branson 1. PLANT Coleoptileanthocyanin Absent Absent Juvenile plant growth Semi-Erect Semi-ErectPlant color at boot stage Green Green Flag leaf at boot stage Erect,twisted Erect, twisted waxy bloom waxy bloom absent present Days toHeading (Julian Date) 136 133 Anther color Yellow Yellow Plant height(cm) 102  97 2. STEM Anthocyanin Absent Present Waxy bloom PresentPresent Internode: Form Hollow Hollow Number  5  5 Hairiness of lastinternode of Absent Present rachis Peduncle: Form Recurved — Length (cm)  14.5   14.0 Auricle: Anthocyanin Absent Present Hair Absent Present 3.HEAD (at maturity) Density Lax Middense Shape Tapering Strap CurvatureErect Erect Awnedness Awnless Awnletted 4. GLUMES (at maturity) ColorWhite White Shoulder Oblique Oblique Shoulder width Narrow Narrow BeakAcute and Obtuse and narrow medium Glume length Long Long Glume widthNarrow Narrow Pubescence Absent Absent 5. SEED Shape Ovate Ovate CheekRounded Rounded Brush Medium Medium Brush collar Not collared Notcollared Crease width Narrow, less Narrow, less than 60% of than 60% ofkernel kernel Crease depth Medium, less Shallow, less than 35% of than20% of kernel kernel Color Red Red Texture Soft Soft Seed weight (g/1000kernels)  39  37 Germ size Midsize Midsize Phenol reaction Fawn Nottested 6. DISEASE REACTIONS Puccinia recondita f. sp. triticiIntermediate Intermediate (Leaf rust): Puccinia striiformis (Striperust): Tolerant Tolerant Stagonospora nodorum (Glume Tolerant Not testedblotch): Septoria tritici Intermediate Tolerant (Speckled leaf blotch):Erysiphe graminis f. sp. tritici Intermediate Tolerant (Powdery mildew):Fusarium spp (Scab): Intermediate Intermediate Soil borne mosaic virus(SBMV): Intermediate Intermediate

In an embodiment, the invention provides a composition comprising a seedof WB-112 comprised in plant seed growth media. Advantageously, plantseed growth media can provide adequate physical support for seeds andcan retain moisture and/or nutritional components. In certainembodiments, the plant seed growth media is a soil or syntheticcultivation medium. Any plant seed growth media known in the art may beutilized in this embodiment and the invention is not limited to soil orsynthetic cultivation medium. Synthetic plant cultivation media are wellknown in the art and may, in certain embodiments, comprise polymers orhydrogels. In specific embodiments, the growth medium may be comprisedin a container or may, for example, be soil in a field.

In another embodiment, the invention is directed to methods forproducing a wheat plant by crossing a first parent wheat plant with asecond parent wheat plant, wherein the first or second wheat plant isthe wheat plant from the cultivar WB-112. In an embodiment, the firstand second parent wheat plants may be from the cultivar WB-112(self-pollination). Any methods using the cultivar WB-112 are part ofthis invention: selfing, backcrosses, hybrid breeding, and crosses topopulations. Any plants produced using cultivar WB-112 as a parent arewithin the scope of this invention. In certain embodiments, theinvention is also directed to cells which, upon growth anddifferentiation, produce a cultivar having essentially all of thephysiological and morphological characteristics of WB-112. The presentinvention additionally contemplates, in various embodiments, a wheatplant regenerated from a tissue culture of cultivar WB-112.

In some embodiments of the invention, the invention is directed to atransgenic variant of WB-112. A transgenic variant of WB-112 may containat least one transgene but could contain at least 1, 2, 3, 4, 5, 6, 7,8, 9, 10, or more transgenes. In another embodiment, a transgenicvariant of WB-112 may contain no more than 15, 14, 13, 12, 11, 10, 9, 8,7, 6, 5, 4, 3, or 2 transgenes. Another embodiment of the inventioninvolves a process for producing wheat variety WB-112 further comprisinga desired trait, said process comprising introducing a transgene thatconfers a desired trait to a wheat plant of variety WB-112. Methods forproducing transgenic plants have been developed and are well known inthe art. As part of the invention, one of ordinary skill in the art mayutilize any method of producing transgenic plants which is currentlyknown or yet to be developed.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available.

In certain embodiments, the desired trait may be one or more ofherbicide tolerance or resistance, insect resistance or tolerance,disease resistance or tolerance, resistance for bacterial, viral, orfungal disease, male fertility, male sterility, decreased phytate, ormodified fatty acid or carbohydrate metabolism. The specific transgenemay be any known in the art or listed herein, including, but not limitedto a polynucleotide conferring resistance to imidazolinone, dicamba,sulfonylurea, glyphosate, glufosinate, triazine, benzonitrile,cyclohexanedione, phenoxy proprionic acid and L-phosphinothricin; apolynucleotide encoding a Bacillus thuringiensis polypeptide, apolynucleotide encoding phytase, FAD-2, FAD-3, galactinol synthase or araffinose synthetic enzyme, Fusarium, Septoria, or various viruses orbacteria.

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include, but are not limited to genes,coding sequences, inducible, constitutive, and tissue specificpromoters, enhancing sequences, and signal and targeting sequences.

In some embodiments, the invention comprises a WB-112 plant which hasbeen developed using both genetic engineering and traditional breedingtechniques. For example, a genetic trait may have been engineered intothe genome of a particular wheat plant may then be moved into the genomeof a WB-112 plant using traditional breeding techniques that are wellknown in the plant breeding arts. Likewise, a genetic trait may havebeen engineered into the genome of a WB-112 wheat plant may then bemoved into the genome of another variety using traditional breedingtechniques that are well known in the plant breeding arts. Abackcrossing approach is commonly used to move a transgene from atransformed wheat variety into an already developed wheat variety, andthe resulting backcross conversion plant would then comprise thetransgene(s).

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid, and can be used alone or incombination with other plasmids, to provide transformed wheat plants,using transformation methods as described below to incorporatetransgenes into the genetic material of the wheat plant(s).

Expression Vectors for Wheat Transformation: Marker Genes

Expression vectors include at least one genetic marker operably linkedto a regulatory element that allows transformed cells containing themarker to be either recovered by negative selection, i.e., inhibitinggrowth of cells that do not contain the selectable marker gene, or bypositive selection, i.e., screening for the product encoded by thegenetic marker. Many commonly used selectable marker genes for planttransformation are well known in the transformation arts, and include,for example, genes that code for enzymes that metabolically detoxify aselective chemical agent which may be an antibiotic or an herbicide, orgenes that encode an altered target which is insensitive to theinhibitor. Positive selection methods are also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII), which when under the control ofplant regulatory signals, confers resistance to kanamycin. Anothercommonly used selectable marker gene is the hygromycinphosphotransferase gene, which confers resistance to the antibiotichygromycin.

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, and aminoglycoside-3′-adenyltransferase, the bleomycin resistance determinant. Other selectablemarker genes confer tolerance or resistance to herbicides such asglyphosate, glufosinate or bromoxynil.

Other selectable marker genes for plant transformation that are not ofbacterial origin include, for example, mouse dihydrofolate reductase,plant 5-enolpyruvyl-shikimate-3-phosphate synthase and plantacetolactate synthase.

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells, rather than directgenetic selection of transformed cells, for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase and chloramphenicol acetyltransferase. Invivo methods for visualizing GUS activity that do not requiredestruction of plant tissue are also available.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. GFP and mutants of GFP may be used as screenable markers.

Expression Vectors for Wheat Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred”.Promoters which initiate transcription only in certain tissue arereferred to as “tissue-specific”. A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may affect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters—An inducible promoter is operably linked to agene for expression in wheat. Optionally, the inducible promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in wheat. With aninducible promoter, the rate of transcription increases in response toan inducing agent.

Any inducible promoter can be used in the present invention. Exemplaryinducible promoters include, but are not limited to, those from the ACEIsystem, which respond to copper; In2 gene from maize, which responds tobenzenesulfonamide herbicide safeners. In an embodiment, the induciblepromoter is a promoter that responds to an inducing agent to whichplants do not normally respond. An exemplary inducible promoter is theinducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone.

B. Constitutive Promoters—A constitutive promoter is operably linked toa gene for expression in wheat or is operably linked to a nucleotidesequence encoding a signal sequence which is operably linked to a genefor expression in wheat.

Many different constitutive promoters can be utilized in the presentinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMVand the promoters from such genes as rice actin; ubiquitin; pEMU; MASand maize H3 histone.

The ALS promoter, Xba1/NcoI fragment 5′ to the Brassica napus ALS3structural gene (or a nucleotide sequence similarity to said Xba1/NcoIfragment), represents a particularly useful constitutive promoter.

C. Tissue-specific or Tissue-preferred Promoters—A tissue-specificpromoter is operably linked to a gene for expression in wheat.Optionally, the tissue-specific promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in wheat. Plants transformed with a gene ofinterest operably linked to a tissue-specific promoter may produce theprotein product of the transgene exclusively, or preferentially, in aspecific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in thepresent invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene; a leaf-specific and light-inducedpromoter, such as that from cab or rubisco; an anther-specific promoter,such as that from LAT52; a pollen-specific promoter, such as that fromZm13; or a microspore-preferred promoter, such as that from apg.

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art.

Foreign Protein Genes and Agronomic Genes

Using transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner. A foreign protein can then beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods.

According to an embodiment of the invention, the transgenic plantprovided for commercial production of foreign protein is, or is derivedfrom a WB-112 wheat plant. In another embodiment, the biomass ofinterest is or is derived from a WB-112 seed. For the relatively smallnumber of transgenic plants that show higher levels of expression, agenetic map can be generated, primarily via conventional restrictionfragment length polymorphism (RFLP), polymerase chain reaction (PCR) andsimple sequence repeat (SSR) analysis, which identifies the approximatechromosomal location of the integrated DNA molecule. Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

In certain embodiments, the invention comprises transformed WB-112plants that express particular agronomic genes or phenotypes ofagronomic interest. Exemplary genes implicated in this regard include,but are not limited to, those categorized below:

1. Genes that Confer Tolerance or Resistance to Pests or Disease andthat Encode:

A. Plant disease tolerance or resistance genes. Plant defenses are oftenactivated by specific interaction between the product of a diseasetolerance or resistance gene (R) in the plant and the product of acorresponding avirulence (Avr) gene in the pathogen. A plant variety canbe transformed with one or more cloned resistance genes to engineerplants that are resistant to specific pathogen strains.

B. A gene conferring resistance to a pest, such as nematodes.

C. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. Moreover, DNA molecules encodingδ-endotoxin genes can be purchased from American Type CultureCollection, Manassas, Va., for example, under ATCC Accession Nos. 40098,67136, 31995 and 31998.

D. A lectin. The nucleotide sequence of several Clivia miniatamannose-binding lectin genes are known in the art.

E. A vitamin-binding protein such as avidin or avidin homologues.

F. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. For example, the nucleotide sequences of ricecysteine proteinase inhibitor, cDNA encoding tobacco proteinaseinhibitor I, Streptomyces nitrosporeus α-amylase inhibitor are known inthe art.

G. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. For example, the baculovirus expressionof cloned juvenile hormone esterase, an inactivator of juvenile hormone,is known in the art.

H. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, it is knownthat expression cloning yields DNA coding for insect diuretic hormonereceptor and an allostatin can be identified in Diploptera puntata.Genes encoding insect-specific, paralytic neurotoxins are also known inthe art.

I. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, heterologous expression in plants of a gene coding for ascorpion insectotoxic peptide is known in the art.

J. An enzyme responsible for a hyper-accumulation of a monoterpene, asesquiterpene, a steroid, a hydroxamic acid, a phenylpropanoidderivative or another non-protein molecule with insecticidal activity.

K. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule. Forexample, such enzymes include, but are not limited to, a glycolyticenzyme, a proteolytic enzyme, a lipolytic enzyme, a nuclease, a cyclase,a callase, a transaminase, an esterase, a hydrolase, a phosphatase, akinase, a phosphorylase, a polymerase, an elastase, a chitinase and aglucanase, whether natural or synthetic. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. The nucleotide sequences of a cDNAencoding tobacco hookworm chitinase and parsley ubi4-2 polyubiquitingene are known.

L. A molecule that stimulates signal transduction. For example, thenucleotide sequences for mung bean calmodulin cDNA clones and a maizecalmodulin cDNA clone are known.

M. A hydrophobic moment peptide. For example, peptide derivatives ofTachyplesin, which inhibit fungal plant pathogens, or syntheticantimicrobial peptides that confer disease resistance.

N. A membrane permease, a channel former, or a channel blocker. Forexample, heterologous expression of a cecropin-β lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearumis known.

O. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmentaffected by the virus from which the coat protein gene is derived, aswell as by related viruses. Coat protein-mediated resistance has beenconferred upon transformed plants against alfalfa mosaic virus, cucumbermosaic virus, tobacco streak virus, potato virus X, potato virus Y,tobacco etch virus, tobacco rattle virus, and tobacco mosaic virus.

P. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Forexample, enzymatic inactivation in transgenic tobacco via production ofsingle-chain antibody fragments is known.

Q. A virus-specific antibody. For example, it is known that transgenicplants expressing recombinant antibody genes are protected from virusattack.

R. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo-α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. The cloning and characterizationof a gene that encodes a bean endopolygalacturonase-inhibiting proteinis known.

S. A developmental-arrestive protein produced in nature by a plant. Forexample, it has been shown that transgenic plants expressing the barleyribosome-inactivating gene have an increased resistance to fungaldisease.

T. Genes expressing proteins with antifungal action. Fusarium headblight along with deoxynivalenol both produced by the pathogen Fusariumgraminearum (Schwabe) have caused devastating losses in wheatproduction. Genes expressing proteins with antifungal action can be usedas transgenes to prevent Fusarium head blight. Various classes ofproteins have been identified. Examples include endochitinases,exochitinases, glucanases, thionins, thaumatin-like proteins, osmotins,ribosome-inactivating proteins, flavonoids and lactoferricin. Duringinfection with Fusarium graminearum, deoxynivalenol is produced. Thereis evidence that production of deoxynivalenol increases the virulence ofthe disease. Genes with properties for detoxification of deoxynivalenolhave been engineered for use in wheat. A synthetic peptide that competeswith deoxynivalenol has been identified. Changing the ribosomes of thehost so that they have reduced affinity for deoxynivalenol has also beenused to reduce the virulence of Fusarium graminearum. Genes used to helpreduce Fusarium head blight include but are not limited to Tri101(Fusarium), PDR5 (yeast), tlp-1 (oat), tlp-2 (oat), leaf tlp-1 (wheat),tlp (rice), tlp-4 (oat), endochitinase, exochitinase, glucanase(Fusarium), permatin (oat), seed hordothionin (barley), alpha-thionin(wheat), acid glucanase (alfalfa), chitinase (barley and rice), classbeta II-1,3-glucanase (barley), PR5/tlp (Arabidopsis), zeamatin (maize),type 1 RIP (barley), NPR1 (Arabidopsis), lactoferrin (mammal),oxalyl-CoA-decarboxylase (bacterium), IAP (baculovirus), ced-9 (C.elegans), and glucanase (rice and barley).

U. A gene, for example, the H9, H10 and H21 genes, conferring resistanceto a pest, such as Hessian fly, stem soft fly, cereal leaf beetle,and/or green bug.

V. A gene conferring resistance to diseases such as wheat rusts,Septoria tritici, Septoria nodorum, powdery mildew, Helminthosporiumdiseases, smuts, bunts, Fusarium diseases, bacterial diseases, and viraldiseases.

W. Genes involved in the Systemic Acquired Resistance (SAR) responseand/or the pathogenesis-related genes.

X. Antifungal genes.

Y. Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.

Z. Cystatin and cysteine proteinase inhibitors.

AA. Defensin genes.

BB. Genes conferring resistance to nematodes.

2. Genes that Confer Tolerance or Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme.

B. Glyphosate (resistance conferred by mutant5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus PAT bar genes), and pyridinoxy or phenoxy proprionic acidsand cyclohexones (ACCase inhibitor-encoding genes). For example, thenucleotide sequence of a form of EPSP which can confer glyphosateresistance is known. A DNA molecule encoding a mutant aroA gene can beobtained under ATCC accession number 39256, and the nucleotide sequenceof the mutant gene is known. Nucleotide sequences of glutaminesynthetase genes which confer tolerance or resistance to herbicides suchas L-phosphinothricin are also known. The nucleotide sequence of a PATgene is known, as is the production of transgenic plants that expresschimeric bar genes coding for PAT activity. Exemplary genes conferringresistance to phenoxy proprionic acids and cyclohexones, such assethoxydim and haloxyfop are the Acc1-S1, Acc1-S2 and Acc1-S3 genes.

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) or a benzonitrile (nitrilase gene). Nucleotide sequencesfor nitrilase genes are disclosed and DNA molecules containing thesegenes are available under ATCC Accession Nos. 53435, 67441, and 67442.Cloning and expression of DNA coding for a glutathione S-transferase isdescribed in the art.

D. Acetohydroxy acid synthase. This enzyme has been found to make plantsthat express this enzyme tolerant or resistant to multiple types ofherbicides and has been introduced into a variety of plants. Other genesthat confer tolerance or resistance to herbicides include a geneencoding a chimeric protein of rat cytochrome P4507A1 and yeastNADPH-cytochrome P450 oxidoreductase, genes for glutathione reductaseand superoxide dismutase, and genes for various phosphotransferases.

E. Protoporphyrinogen oxidase (protox). Protox is necessary for theproduction of chlorophyll, which is necessary for all plant survival.The protox enzyme serves as the target for a variety of herbicidalcompounds. These herbicides also inhibit growth of different species ofplants present, causing their total destruction. The development ofplants containing altered protox activity that are tolerant or resistantto these herbicides is described in the art.

3. Genes that Confer or Contribute to a Value-Added Trait, such as:

A. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant.

B. Decreased phytate content. This can be accomplished by: 1)Introduction of a phytase-encoding gene enhances breakdown of phytate,adding more free phosphate to the transformed plant; or 2) Up-regulationof a gene that reduces phytate content. For example, the nucleotidesequence of an Aspergillus niger phytase gene has been described

C. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch, or, a gene altering thioredoxin such as NTRand/or TRX and/or a gamma zein knock out or mutant such as cs27 orTUSC27 or en27. For example, the nucleotide sequences of Streptococcusmutans fructosyltransferase gene, Bacillus subtilis levansucrase gene,tomato invertase genes are known. Transgenic plants can be produced thatexpress Bacillus licheniformis alpha-amylase, that site-directmutagenesis of barley alpha-amylase gene, or confer maize endospermstarch branching enzyme II or improved digestibility and/or starchextraction through modification of UDP-D-xylose 4-epimerase. Methods ofproducing high oil seed by modification of starch levels (AGP) are alsoknown. The fatty acid modification genes mentioned above may also beused to affect starch content and/or composition through theinterrelationship of the starch and oil pathways.

D. Elevated oleic acid via FAD-2 gene modification and/or decreasedlinolenic acid via FAD-3 gene modification.

E. Altering conjugated linolenic or linoleic acid content, or LEC1, AGP,Dek1, Superal1, mi1ps, various Ipa genes such as Ipa1, Ipa3, hpt orhggt.

F. Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, manipulation of antioxidantlevels through alteration of a phytl prenyl transferase (ppt) is known,as is manipulation of antioxidant levels through alteration of ahomogentisate geranyl geranyl transferase (hggt).

G. The content of high-molecular weight gluten subunits (HMS-GS).Genomic clones have been isolated for different subunits. For example,genomic clones have transformed wheat with genes that encode a modifiedHMW-GS.

H. Increased protein metabolism, zinc and iron content, for example, byregulating the NAC gene, increasing protein metabolism by regulating theGpc-B1 gene, or regulating glutenin and gliadin genes.

I. Altered essential seed amino acids. Methods of increasingaccumulation of essential amino acids in seeds are known, binary methodsof increasing accumulation of essential amino acids in seeds are known,alteration of amino acid compositions in seeds are known, methods foraltering amino acid content of proteins are known, alteration of aminoacid compositions in seeds are known, and proteins with enhanced levelsof essential amino acids are known. Other examples may include highmethionine, high threonine, plant amino acid biosynthetic enzymes,increased lysine and threonine, plant tryptophan synthase beta subunit,methionine metabolic enzymes, high sulfur, increased methionine, plantamino acid biosynthetic enzymes, engineered seed protein having higherpercentage of essential amino acids, increased lysine, increasing sulfuramino acid content, synthetic storage proteins with defined structurecontaining programmable levels of essential amino acids for improvementof the nutritional value of plants, increased threonine, increasedlysine, Ces A: cellulose synthase, hemicellulose, UDPGdH, and RGP.

4. Genes that Control Male Sterility

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility. In addition to these methods, asystem of nuclear male sterility that includes: identifying a gene whichis critical to male fertility; silencing this native gene which iscritical to male fertility; removing the native promoter from theessential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on”, resulting in the male fertility gene notbeing transcribed, is known. Fertility is restored by inducing, orturning “on”, the promoter, which in turn allows the gene that confersmale fertility to be transcribed.

A. Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN—Ac-PPT.

B. Introduction of various stamen-specific promoters.

C. Introduction of the barnase and the barstar genes.

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.Other systems that may be used include the Gin recombinase of phage Mu,the Pin recombinase of E. coli, and the R/RS system of the pSR1 plasmid.

6. Genes that Affect Abiotic Stress Resistance.

A. Genes that affect abiotic stress resistance (including but notlimited to flowering, seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress. For example,water use efficiency can be altered through alteration of malate. Inaddition, various genes, including CBF genes and transcription factorscan be effective in mitigating the negative effects of freezing, highsalinity, and drought on plants, as well as conferring other positiveeffects on plant phenotype. Abscisic acid can be altered in plants,resulting in improved plant phenotype, such as increased yield and/orincreased tolerance to abiotic stress. Cytokinin expression can bemodified resulting in plants with increased stress tolerance, such asdrought tolerance, and/or increased yield. Nitrogen utilization can beenhanced and/or nitrogen responsiveness can be altered. Ethylene can bealtered. Plant transcription factors or transcriptional regulators ofabiotic stress can also be altered.

B. Improved tolerance to water stress from drought or high salt watercondition. The HVA1 protein belongs to the group 3 LEA proteins thatinclude other members such as wheat pMA2005, cotton D-7, carrot Dc3, andrape pLEA76. These proteins are characterized by 11-mer tandem repeatsof amino acid domains which may form a probable amphophilicalpha-helical structure that presents a hydrophilic surface with ahydrophobic stripe. The barley HVA1 gene and the wheat pMA2005 gene arehighly similar at both the nucleotide level and predicted amino acidlevel. These two monocot genes are closely related to the cotton D-7gene and carrot Dc3 gene with which they share a similar structural geneorganization. There is, therefore, a correlation between LEA geneexpression or LEA protein accumulation with stress tolerance in a numberof plants. For example, in severely dehydrated wheat seedlings, theaccumulation of high levels of group 3 LEA proteins was correlated withtissue dehydration tolerance. Studies on several Indica varieties ofrice showed that the levels of group 2 LEA proteins (also known asdehydrins) and group 3 LEA proteins in roots were significantly higherin salt-tolerant varieties compared with sensitive varieties. The barleyHVA1 gene was transformed into wheat. Transformed wheat plants showedincreased tolerance to water stress.

C. Improved water stress tolerance through increased mannitol levels viathe bacterial mannitol-1-phosphate dehydrogenase gene. It is known toproduce a plant with a genetic basis for coping with water deficit byintroduction of the bacterial mannitol-1-phosphate dehydrogenase gene,mtlD, into tobacco cells via Agrobacterium-mediated transformation. Rootand leaf tissues from transgenic plants regenerated from thesetransformed tobacco cells contained up to 100 mM mannitol. Controlplants contained no detectable mannitol. To determine whether thetransgenic tobacco plants exhibited increased tolerance to waterdeficit, the growth of transgenic plants was compared to that ofuntransformed control plants in the presence of 250 mM NaCl. After 30days of exposure to 250 mM NaCl, transgenic plants had decreased weightloss and increased height relative to their untransformed counterparts.The authors concluded that the presence of mannitol in these transformedtobacco plants contributed to water deficit tolerance at the cellularlevel.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants.

Methods for Wheat Transformation

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available.

A. Agrobacterium-Mediated Transformation. One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. A. tumefaciens and A. rhizogenes are plantpathogenic soil bacteria which genetically transform plant cells. The Tiand Ri plasmids of A. tumefaciens and A. rhizogenes, respectively, carrygenes responsible for genetic transformation of the plant. Descriptionsof Agrobacterium vector systems and methods for Agrobacterium-mediatedgene transfer are known.

B. Direct Gene Transfer. Several methods of plant transformation,collectively referred to as direct gene transfer, have been developed asan alternative to Agrobacterium-mediated transformation. A generallyapplicable method of plant transformation is microprojectile-mediatedtransformation wherein DNA is carried on the surface of microprojectilesmeasuring 1 to 4 μm. The expression vector is introduced into planttissues with a biolistic device that accelerates the microprojectiles tospeeds of 300 to 600 m/s which is sufficient to penetrate plant cellwalls and membranes.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Alternatively, liposome and spheroplast fusion have beenused to introduce expression vectors into plants. Direct uptake of DNAinto protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine has also been reported. Electroporation of protoplastsand whole cells and tissues has also been described. Followingtransformation of wheat target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed, with another (non-transformed or transformed) variety, in orderto produce a new transgenic variety. Alternatively, a genetic traitwhich has been engineered into a particular wheat cultivar using theforegoing transformation techniques could be moved into another cultivarusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties which do not contain that gene. As usedherein, “crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context.

Genetic Marker Profile Through SSR and First Generation Progeny

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile which can identify plants of the same variety ora related variety or be used to determine or validate a pedigree.Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asMicrosatellites, and Single Nucleotide Polymorphisms (SNPs).

Particular markers used for these purposes are not limited to anyparticular set of markers, but are envisioned to include any type ofmarker and marker profile which provides a means of distinguishingvarieties. One method of comparison is to use only homozygous loci forWB-112.

In addition to being used for identification of wheat variety WB-112 andplant parts and plant cells of variety WB-112, the genetic profile maybe used to identify a wheat plant produced through the use of WB-112 orto verify a pedigree for progeny plants produced through the use ofWB-112. The genetic marker profile is also useful in breeding anddeveloping backcross conversions.

In some embodiments, the present invention comprises a wheat plantcharacterized by molecular and physiological data obtained from therepresentative sample of WB-112, deposited with the American TypeCulture Collection (ATCC). Provided in further embodiments of theinvention is a wheat plant formed by the combination of the WB-112 plantor plant cell with another wheat plant or cell and comprising thehomozygous alleles of the variety.

Means of performing genetic marker profiles using SSR polymorphisms arewell known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. A markersystem based on SSRs can be highly informative in linkage analysisrelative to other marker systems in that multiple alleles may bepresent. Another advantage of this type of marker is that, through useof flanking primers, detection of SSRs can be achieved, for example, bythe polymerase chain reaction (PCR), thereby eliminating the need forlabor-intensive Southern hybridization. PCR detection uses twooligonucleotide primers flanking the polymorphic segment of repetitiveDNA. Repeated cycles of heat denaturation of the DNA, followed byannealing of the primers to their complementary sequences at lowtemperatures, and extension of the annealed primers with DNA polymerase,comprise the major part of the methodology.

Following amplification, markers can be scored by electrophoresis of theamplification products. Scoring of marker genotype is based on the sizeof the amplified fragment, which may be measured by the number of basepairs of the fragment. While variation in the primer used or inlaboratory procedures can affect the reported fragment size, relativevalues should remain constant regardless of the specific primer orlaboratory used. When comparing varieties, all SSR profiles may beperformed in the same lab.

The SSR profile of wheat plant WB-112 can be used to identify plantscomprising WB-112 as a parent, since such plants will comprise the samehomozygous alleles as WB-112. Because the wheat variety is essentiallyhomozygous at all relevant loci, most loci should have only one type ofallele present. In contrast, a genetic marker profile of an F₁ progenyshould be the sum of those parents, e.g., if one parent was homozygousfor allele x at a particular locus, and the other parent homozygous forallele y at that locus, then the F₁ progeny will be xy (heterozygous) atthat locus. Subsequent generations of progeny produced by selection andbreeding are expected to be of genotype x (homozygous), y (homozygous),or xy (heterozygous) for that locus position. When the F₁ plant isselfed or sibbed for successive filial generations, the locus should beeither x or y for that position.

In addition, plants and plant parts substantially benefiting from theuse of WB-112 in their development, such as WB-112 comprising abackcross conversion, transgene, or genetic sterility factor, may beidentified by having a molecular marker profile with a high percentidentity to WB-112. In an embodiment, such a percent identity might be95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to WB-112.

The SSR profile of WB-112 also can be used to identify essentiallyderived varieties and other progeny varieties developed from the use ofWB-112, as well as cells and other plant parts thereof. Progeny plantsand plant parts produced using WB-112 may be identified by having amolecular marker profile of at least 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or99.5% genetic contribution from WB-112, as measured by either percentidentity or percent similarity. Such progeny may be furthercharacterized as being within a pedigree distance of WB-112, such aswithin 1, 2, 3, 4 or 5 or fewer cross-pollinations to a wheat plantother than WB-112 or a plant that has WB-112 as a progenitor. Uniquemolecular profiles may be identified with other molecular tools such asSNPs and RFLPs.

While determining the SSR genetic marker profile of the plants describedsupra, several unique SSR profiles may also be identified which did notappear in either parent of such plant. Such unique SSR profiles mayarise during the breeding process from recombination or mutation. Acombination of several unique alleles provides a means of identifying aplant variety, an F₁ progeny produced from such variety, and furtherprogeny produced from such variety.

Gene Conversion

When the term “wheat plant” is used in the context of the presentinvention, this also includes any gene conversions of that variety.Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the variety. For example, a varietymay be backcrossed 1, 2, 3, 4, 5, 6, 7, 8, 9 or more times to therecurrent parent. The parental wheat plant which contributes the genefor the desired characteristic is termed the “nonrecurrent” or “donorparent”. This terminology refers to the fact that the nonrecurrentparent is used one time in the backcross protocol and therefore does notrecur. The parental wheat plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol. In a typicalbackcross protocol, the original variety of interest (recurrent parent)is crossed to a second variety (nonrecurrent parent) that carries thesingle gene of interest to be transferred. The resulting progeny fromthis cross are then crossed again to the recurrent parent and theprocess is repeated until a wheat plant is obtained wherein essentiallyall of the desired morphological and physiological characteristics ofthe recurrent parent are recovered in the converted plant, in additionto the single transferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent contributes to a successfulbackcrossing procedure. The goal of a backcross protocol is to alter orsubstitute a single trait or characteristic in the original variety. Toaccomplish this, a single gene of the recurrent variety is modified orsubstituted with the desired gene from the nonrecurrent parent, whileretaining essentially all of the rest of the desired genetic, andtherefore the desired physiological and morphological, constitution ofthe original variety. The choice of the particular nonrecurrent parentwill depend on the purpose of the backcross. One of the major purposesis to add commercially desirable, agronomically important traits to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele may also be transferred. In this instance, it may benecessary to introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety, but that can beimproved by backcrossing techniques. Single gene traits may or may notbe transgenic. Examples of these traits include, but are not limited to,male sterility, waxy starch, herbicide tolerance or resistance,resistance for bacterial, fungal, or viral disease, insect resistance ortolerance, male fertility, enhanced nutritional quality, industrialusage, yield stability and yield enhancement. These genes are generallyinherited through the nucleus.

Introduction of a New Trait or Locus into WB-112

Variety WB-112 represents a new base genetic variety into which a newlocus or trait may be introgressed. Direct transformation andbackcrossing represent two important methods that can be used toaccomplish such an introgression. The term backcross conversion andsingle locus conversion are used interchangeably to designate theproduct of a backcrossing program.

Backcross Conversions of WB-112

A backcross conversion of WB-112 occurs when DNA sequences areintroduced through backcrossing, with WB-112 utilized as the recurrentparent. Both naturally occurring and transgenic DNA sequences may beintroduced through backcrossing techniques. A backcross conversion mayproduce a plant with a trait or locus conversion in at least two or morebackcrosses, including at least 2 crosses, at least 3 crosses, at least4 crosses, at least 5 crosses, or additional crosses. Molecular markerassisted breeding or selection may be utilized to reduce the number ofbackcrosses necessary to achieve the backcross conversion. For example,a backcross conversion can be made in as few as two backcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes versusunlinked genes), the level of expression of the trait, the type ofinheritance (cytoplasmic or nuclear) and the types of parents includedin the cross. It is understood by those of ordinary skill in the artthat for single gene traits that are relatively easy to classify, thebackcross method is effective and relatively easy to manage. Desiredtraits that may be transferred through backcross conversion include, butare not limited to, sterility (nuclear and cytoplasmic), fertilityrestoration, nutritional enhancements, drought tolerance, nitrogenutilization, altered fatty acid profile, low phytate, industrialenhancements, disease resistance or tolerance (bacterial, fungal orviral), insect resistance or tolerance, and herbicide tolerance orresistance. In addition, an introgression site itself, such as an FRTsite, Lox site or other site specific integration site, may be insertedby backcrossing and utilized for direct insertion of one or more genesof interest into a specific plant variety. In some embodiments of theinvention, the number of loci that may be backcrossed into WB-112 is atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. A single locus may containseveral transgenes, such as a transgene for disease resistance that, inthe same expression vector, also contains a transgene for herbicidetolerance or resistance. The gene for herbicide tolerance or resistancemay be used as a selectable marker and/or as a phenotypic trait. Asingle locus conversion of site specific integration system allows forthe integration of multiple genes at the converted loci.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selfing thefirst backcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the locusof interest. The last backcross generation is usually selfed to givepure breeding progeny for the gene(s) being transferred, although abackcross conversion with a stably introgressed trait may also bemaintained by further backcrossing to the recurrent parent withselection for the converted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype of the recurrent parent. The backcross is a form ofinbreeding, and the features of the recurrent parent are automaticallyrecovered after successive backcrosses. Some sources suggest from one tofour or more backcrosses, but as noted above, the number of backcrossesnecessary can be reduced with the use of molecular markers. Otherfactors, such as a genetically similar donor parent, may also reduce thenumber of backcrosses necessary. Backcrossing is easiest for simplyinherited, dominant and easily recognized traits.

One process for adding or modifying a trait or locus in wheat varietyWB-112 comprises crossing WB-112 plants grown from WB-112 seed withplants of another wheat variety that comprise the desired trait orlocus, selecting F₁ progeny plants that comprise the desired trait orlocus to produce selected F₁ progeny plants, crossing the selectedprogeny plants with the WB-112 plants to produce backcross progenyplants, selecting for backcross progeny plants that have the desiredtrait or locus and the morphological characteristics of wheat varietyWB-112 to produce selected backcross progeny plants, and backcrossing toWB-112 three or more times in succession to produce selected fourth orhigher backcross progeny plants that comprise said trait or locus. Themodified WB-112 may be further characterized as having essentially allof the physiological and morphological characteristics of wheat varietyWB-112 listed in Table 1, as determined at the 5% significance levelwhen grown in the same environmental conditions and/or may becharacterized by percent similarity or identity to WB-112 as determinedby SSR markers. The above method may be utilized with fewer backcrossesin appropriate situations, such as when the donor parent is highlyrelated or markers are used in the selection step. Desired nucleic acidsthat may be used include those nucleic acids known in the art, some ofwhich are listed herein, that will affect traits through nucleic acidexpression or inhibition. Desired loci include the introgression of FRT,Lox and other sites for site specific integration, which may also affecta desired trait if a functional nucleic acid is inserted at theintegration site.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny wheat seed byadding a step at the end of the process that comprises crossing WB-112with the introgressed trait or locus with a different wheat plant andharvesting the resultant first generation progeny wheat seed.

A further embodiment of the invention is a backcross conversion of wheatvariety WB-112. A backcross conversion occurs when DNA sequences areintroduced through traditional (non-transformation) breeding techniques,such as backcrossing. DNA sequences, whether naturally occurring ortransgenes, may be introduced using these traditional breedingtechniques. Desired traits transferred through this process include, butare not limited to nutritional enhancements, industrial enhancements,disease resistance or tolerance, insect resistance or tolerance,herbicide tolerance or resistance, agronomic enhancements, grain qualityenhancement, waxy starch, breeding enhancements, seed productionenhancements, and male sterility. Descriptions of some of thecytoplasmic male sterility genes, nuclear male sterility genes, chemicalhybridizing agents, male fertility restoration genes, and methods ofusing the aforementioned are known. Examples of genes for other traitsinclude: Leaf rust resistance genes (Lr series such as Lr1, Lr10, Lr21,Lr22, Lr22a, Lr32, Lr37, Lr41, Lr42, and Lr43), Fusarium headblight-resistance genes (QFhs.ndsu-3B and QFhs.ndsu-2A), powdery mildewresistance genes (Pm21), common bunt resistance genes (Bt-10), and wheatstreak mosaic virus resistance gene (Wsm1), Russian wheat aphidresistance genes (Dn series such as Dn1, Dn2, Dn4, Dn5), Black stem rustresistance genes (Sr38), Yellow rust resistance genes (Yr series such asYr1, YrSD, Yrsu, Yr17, Yr15, YrH52), Aluminum tolerance genes (Alt(BH)),dwarf genes (Rht), vernalization genes (Vrn), Hessian fly resistancegenes (H9, H10, H21, H29), grain color genes (R/r), glyphosateresistance genes (EPSPS), glufosinate genes (bar, pat) and water stresstolerance genes (Hva1, mtlD). The trait of interest is transferred fromthe donor parent to the recurrent parent, which in this case, is thewheat plant disclosed herein, WB-112. Single gene traits may result fromeither the transfer of a dominant allele or a recessive allele.Selection of progeny containing the trait of interest is done by directselection for a trait associated with a dominant allele. Selection ofprogeny for a trait that is transferred via a recessive allele requiresgrowing and selfing the first backcross to determine which plants carrythe recessive alleles. Recessive traits may require additional progenytesting in successive backcross generations to determine the presence ofthe gene of interest.

Using WB-112 to Develop Other Wheat Varieties

Wheat varieties such as WB-112 are typically developed for use in seedand grain production. However, wheat varieties such as WB-112 alsoprovide a source of breeding material that may be used to develop newwheat varieties. Plant breeding techniques known in the art and used ina wheat plant breeding program include, but are not limited to,recurrent selection, mass selection, bulk selection, mass selection,backcrossing, pedigree breeding, open pollination breeding, restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, and transformation. Often,combinations of these techniques are used. The development of wheatvarieties in a plant breeding program requires, in general, thedevelopment and evaluation of homozygous varieties. There are manyanalytical methods available to evaluate a new variety. The oldest andmost traditional method of analysis is the observation of phenotypictraits but genotypic analysis may also be used.

Additional Breeding Methods

In an embodiment, this invention is directed to methods for producing awheat plant by crossing a first parent wheat plant with a second parentwheat plant wherein either the first or second parent wheat plant isvariety WB-112. The other parent may be any other wheat plant, such as awheat plant that is part of a synthetic or natural population. Any suchmethods using wheat variety WB-112 are part of this invention: selfing,sibbing, backcrosses, mass selection, pedigree breeding, bulk selection,hybrid production, and crosses to populations. These methods are wellknown in the art and some of the more commonly used breeding methods aredescribed below.

The following describes breeding methods that may be used with wheatcultivar WB-112 in the development of further wheat plants. One suchembodiment is a method for developing a cultivar WB-112 progeny wheatplant in a wheat plant breeding program comprising: obtaining the wheatplant, or a part thereof, of cultivar WB-112 utilizing said plant orplant part as a source of breeding material and selecting a wheatcultivar WB-112 progeny plant with molecular markers in common withcultivar WB-112 and/or with morphological and/or physiologicalcharacteristics selected from the characteristics listed in Tables 1 or2. Breeding steps that may be used in the wheat plant breeding programinclude pedigree breeding, backcrossing, mutation breeding, andrecurrent selection. In conjunction with these steps, techniques such asRFLP-enhanced selection, genetic marker enhanced selection (for exampleSSR markers) and the making of double haploids may be utilized.

Another method involves producing a population of wheat cultivar WB-112progeny wheat plants, comprising crossing cultivar WB-112 with anotherwheat plant, thereby producing a population of wheat plants, which, onaverage, derive 50% of their alleles from wheat cultivar WB-112. A plantof this population may be selected and repeatedly selfed or sibbed witha wheat cultivar resulting from these successive filial generations. Oneembodiment of this invention is the wheat cultivar produced by thismethod and that has obtained at least 50% of its alleles from wheatcultivar WB-112.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. Thus the invention includes wheat cultivar WB-112 progenywheat plants comprising a combination of at least two cultivar WB-112traits selected from the group consisting of those listed in Tables 1and 2, so that said progeny wheat plant is not significantly differentfor said traits than wheat cultivar WB-112. Using techniques describedherein, molecular markers may be used to identify said progeny plant asa wheat cultivar WB-112 progeny plant. Mean trait values may be used todetermine whether trait differences are significant, and the traits maybe measured on plants grown under the same environmental conditions.Once such a variety is developed its value is substantial, as it isimportant to advance the germplasm base as a whole in order to maintainor improve traits such as yield, disease resistance or tolerance, pestresistance or tolerance, and plant performance in extreme environmentalconditions.

Progeny of wheat cultivar WB-112 may also be characterized through theirfilial relationship with wheat cultivar WB-112, as for example, beingwithin a certain number of breeding crosses of wheat cultivar WB-112. Abreeding cross is a cross made to introduce new genetics into theprogeny, and is distinguished from a cross, such as a self or a sibcross, made to select among existing genetic alleles. The lower thenumber of breeding crosses in the pedigree, the closer the relationshipbetween wheat cultivar WB-112 and its progeny. For example, progenyproduced by the methods described herein may be within 1, 2, 3, 4 or 5breeding crosses of wheat cultivar WB-112.

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such asWB-112 and another wheat variety having one or more desirablecharacteristics that is lacking or which complements WB-112. If the twooriginal parents do not provide all the desired characteristics, othersources can be included in the breeding population. In the pedigreemethod, superior plants are selfed and selected in successive filialgenerations. In the succeeding filial generations the heterozygouscondition gives way to homogeneous varieties as a result ofself-pollination and selection. Typically in the pedigree method ofbreeding, five or more successive filial generations of selfing andselection is practiced: F₁ to F₂; F₂ to F₃; F₃ to F₄; F₄ to F₅, etc.After a sufficient amount of inbreeding, successive filial generationswill serve to increase seed of the developed variety. In an embodiment,the developed variety comprises homozygous alleles at about 95% or moreof its loci.

In addition to being used to create a backcross conversion, backcrossingcan also be used in combination with pedigree breeding. As discussedpreviously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety, the donor parent, to adeveloped variety called the recurrent parent, which has overall goodagronomic characteristics yet lacks that desirable trait or traits.However, the same procedure can be used to move the progeny toward thegenotype of the recurrent parent but at the same time retain manycomponents of the non-recurrent parent by stopping the backcrossing atan early stage and proceeding with selfing and selection. For example, awheat variety may be crossed with another variety to produce a firstgeneration progeny plant. The first generation progeny plant may then bebackcrossed to one of its parent varieties to create a BC1 or BC2.Progeny are selfed and selected so that the newly developed variety hasmany of the attributes of the recurrent parent and yet several of thedesired attributes of the non-recurrent parent. This approach leveragesthe value and strengths of the recurrent parent for use in new wheatvarieties.

Therefore, an embodiment of this invention is a method of making abackcross conversion of wheat variety WB-112 comprising the steps ofcrossing a plant of wheat variety WB-112 with a donor plant comprising adesired trait, selecting an F₁ progeny plant comprising the desiredtrait, and backcrossing the selected F₁ progeny plant to a plant ofwheat variety WB-112. This method may further comprise the step ofobtaining a molecular marker profile of wheat variety WB-112 and usingthe molecular marker profile to select for a progeny plant with thedesired trait and the molecular marker profile of WB-112. In oneembodiment, the desired trait is a mutant gene or transgene present inthe donor parent.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. WB-112 is suitable for use in arecurrent selection program. The method entails individual plants crosspollinating with each other to form progeny. The progeny are grown andthe superior progeny selected by any number of selection methods, whichinclude individual plant, half-sib progeny, full-sib progeny and selfedprogeny. The selected progeny are cross pollinated with each other toform progeny for another population. This population is planted andagain superior plants are selected to cross pollinate with each other.Recurrent selection is a cyclical process and therefore can be repeatedas many times as desired. The objective of recurrent selection is toimprove the traits of a population. The improved population can then beused as a source of breeding material to obtain new varieties forcommercial or breeding use, including the production of a syntheticcultivar. A synthetic cultivar is the resultant progeny formed by theintercrossing of several selected varieties.

Mass selection is a useful technique when used in conjunction withmolecular marker enhanced selection. In mass selection, seeds fromindividuals are selected based on phenotype or genotype. These selectedseeds are then bulked and used to grow the next generation. Bulkselection requires growing a population of plants in a bulk plot,allowing the plants to self-pollinate, harvesting the seed in bulk andthen using a sample of the seed harvested in bulk to plant the nextgeneration. Also, instead of self pollination, directed pollinationcould be used as part of the breeding program.

Mutation Breeding

Mutation breeding is another method of introducing new traits into wheatvariety WB-112. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation; such as X-rays, Gamma rays (e.g. cobalt60 or cesium 137), neutrons, (product of nuclear fission by uranium 235in an atomic reactor), Beta radiation (emitted from radioisotopes suchas phosphorus 32 or carbon 14), or ultraviolet radiation (optionallyfrom 2500 to 2900 nm), or chemical mutagens (such as base analogues(5-bromo-uracil), related compounds (8-ethoxy caffeine), antibiotics(streptonigrin), alkylating agents (sulfur mustards, nitrogen mustards,epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones),azide, hydroxylamine, nitrous acid, or acridines. Once a desired traitis observed through mutagenesis the trait may then be incorporated intoexisting germplasm by traditional breeding techniques. In addition,mutations created in other wheat plants may be used to produce abackcross conversion of wheat cultivar WB-112 that comprises suchmutation. Further embodiments of the invention are the treatment ofWB-112 with a mutagen and the plant produced by mutagenesis of WB-112.

Breeding with Molecular Markers

Molecular markers, which includes markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs) and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods utilizing wheat cultivar WB-112. IsozymeElectrophoresis and RFLPs have been widely used to determine geneticcomposition.

SSR technology is currently the most efficient and practical markertechnology; more marker loci can be routinely used and more alleles permarker locus can be found using SSRs in comparison to RFLPs. SingleNucleotide Polymorphisms (SNPs) may also be used to identify the uniquegenetic composition of the invention and progeny varieties retainingthat unique genetic composition. Various molecular marker techniques maybe used in combination to enhance overall resolution. Wheat DNAmolecular marker linkage maps have been rapidly constructed and widelyimplemented in genetic studies.

One use of molecular markers is QTL mapping. QTL mapping is the use ofmarkers which are known to be closely linked to alleles that havemeasurable effects on a quantitative trait. Selection in the breedingprocess is based upon the accumulation of markers linked to the positiveeffecting alleles and/or the elimination of the markers linked to thenegative effecting alleles from the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants. Itcan also be used to reduce the number of crosses back to the recurrentparent needed in a backcrossing program. The use of molecular markers inthe selection process is often called genetic marker enhanced selection.Molecular markers may also be used to identify and exclude certainsources of germplasm as parental varieties or ancestors of a plant byproviding a means of tracking genetic profiles through crosses.

Production of Double Haploids

The production of double haploids can also be used for the developmentof plants with a homozygous phenotype in the breeding program. Forexample, a wheat plant for which wheat cultivar WB-112 is a parent canbe used to produce double haploid plants. Double haploids are producedby the doubling of a set of chromosomes (1 N) from a heterozygous plantto produce a completely homozygous individual. This can be advantageousbecause the process omits the generations of selfing needed to obtain ahomozygous plant from a heterozygous source.

Haploid induction systems have been developed for various plants toproduce haploid tissues, plants and seeds. The haploid induction systemcan produce haploid plants from any genotype by crossing a selected line(as female) with an inducer line. Such inducer lines for maize includeStock 6. Methods for obtaining haploid plants have also been disclosedin the art.

Thus, an embodiment of this invention is a process for making asubstantially homozygous WB-112 progeny plant by producing or obtaininga seed from the cross of WB-112 and another wheat plant and applyingdouble haploid methods to the F₁ seed or F₁ plant or to any successivefilial generation. Based on studies in maize and currently beingconducted in wheat, such methods would decrease the number ofgenerations required to produce a variety with similar genetics orcharacteristics to WB-112.

In particular, a process of making seed retaining the molecular markerprofile of wheat variety WB-112 is contemplated, such process comprisingobtaining or producing F₁ seed for which wheat variety WB-112 is aparent, inducing doubled haploids to create progeny without theoccurrence of meiotic segregation, obtaining the molecular markerprofile of wheat variety WB-112, and selecting progeny that retain themolecular marker profile of WB-112. Descriptions of other breedingmethods that are commonly used for different traits and crops are known.

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of wheat andregeneration of plants therefrom is well known and widely published.Thus, another aspect of this invention is to provide cells which upongrowth and differentiation produce wheat plants having essentially allof the physiological and morphological characteristics of wheat cultivarWB-112. Means for preparing and maintaining plant tissue culture arewell known in the art. By way of example, a tissue culture comprisingorgans has been used to produce regenerated plants.

In Table 2, selected yield, quality and agronomic characteristics ofwheat cultivar WB-112 are compared to three commercial check varieties.Column 1 shows the variety, column 2 shows the yield as a percent of theaverage of the three check varieties, column 3 shows the test weight ofharvested grain, column 4 shows the Julian flowering date when 50% ofthe variety flowers, column 5 shows the plant height in centimeters,column 6 shows the protein level of the harvest grain as a percent(as-is basis) of total grain weight, column 7 shows the Baking Qualityscore, and column 8 shows the Milling Quality score.

TABLE 2 Some Yield and Quality Characteristics of Variety “WB-112”Compared to “Branson”, “P25R47” and “P25R56” 3 4 5 7 8 2 Test FloweringPlant 6 Baking Milling 1 Yield Weight Date Height Protein Quality Score¹Quality Score² Unit of Measure % of Av lbs/bu Julian cm % See FootnoteSee Footnote Branson 103.1 56.2 129.2 94.4 — — — P25R47 109.4 55.0 132.990.6 9.77 74.1 67.3 P25R56 101.4 55.0 132.6 91.2 — — — Average of 100.055.6 131.6 92.1 Checks WB-112 107.6 57.5 132.8 98.8 9.36 74.4 75.1 No.of 79 70 6 6 1 1 1 Replications ¹Baking Quality Score (BQS) BQS = BF +(33.3333 * CS) − 526.667 BF = Allis Baking Score − SCS CS = Cookie Score= (−0.145 * Flour Protein) + (−0.07 * Sucrose SRC) + (0.049 * SE) + 21.9SCS = Standard Cookie Score—cookie score for the quality standarddesignated for the trial as measured in the trial being evaluated AllisBaking Score = Allis baking score for the quality standard as determinedin the Allis Milling Database ²Milling Quality Score (MQS) MQS = MF +(5.0144 * Adjusted Flour Yield) − 292.6425 MF = Allis Milling Score −(5.0144 * SAFY) − 292.6425 Allis Milling Score = Mill score from Allisdatabase for the quality standard designated for the group SAFY =Adjusted Flour Yield for the quality standard designated for the trialas measured in the trial being evaluated

As shown in Table 2, the wheat cultivar of the present invention,WB-112, had a yield that was in between the check varieties, a testweight that was greater than the average of the check varieties, a plantheight that was taller than the average of the check varieties, greaternumber of days to flowering than check varieties, and a baking andmilling quality score yield that was greater than the average of thechecks.

Deposit Information

A deposit of the Monsanto Technology, LLC proprietary common wheatvariety WB-112 disclosed above and recited in the appended claims hasbeen made with the American Type Culture Collection (ATCC), 10801University Boulevard, Manassas, Va. 20110. The date of deposit was Jun.21, 2011. The deposit of 2,500 seeds was taken from the same depositmaintained by Monsanto Technology, LLC since prior to the filing date ofthis application. All restrictions upon the deposit have beenirrevocably removed, and the deposit is intended to meet all of therequirements of 37 C.F.R. 1.801-1.809. The ATCC accession number isPTA-11955. The deposit will be maintained in the depository for a periodof 30 years, or 5 years after the last request, or for the enforceablelife of the patent, whichever is longer, and will be replaced asnecessary during that period.

All references cited in this specification, including withoutlimitation, all papers, publications, patents, patent applications,presentations, texts, reports, manuscripts, brochures, books, internetpostings, journal articles, and/or periodicals are hereby incorporatedby reference into this specification in their entireties. The discussionof the references herein is intended merely to summarize the assertionsmade by their authors and no admission is made that any referenceconstitutes prior art. Applicants reserve the right to challenge theaccuracy and pertinence of the cited references.

These and other modifications and variations to the present inventionmay be practiced by those of ordinary skill in the art, withoutdeparting from the spirit and scope of the present invention, which ismore particularly set forth in the appended claims. In addition, itshould be understood that aspects of the various embodiments may beinterchanged in whole or in part. Furthermore, those of ordinary skillin the art will appreciate that the foregoing description is by way ofexample only, and is not intended to limit the invention so furtherdescribed in such appended claims. Therefore, the spirit and scope ofthe appended claims should not be limited to the description of theversions contained therein.

What is claimed is:
 1. A seed of wheat cultivar WB-112, a representativesample of seed of which was deposited under ATCC Accession No.PTA-11955.
 2. A wheat plant, or a part thereof, produced by growing theseed of claim
 1. 3. A tissue culture produced from protoplasts or cellsfrom the plant of claim
 2. 4. The tissue culture of claim 3, whereinsaid cells or protoplasts are produced from a plant part selected fromthe group consisting of head, awn, leaf, pollen, ovule, embryo,cotyledon, hypocotyl, meristematic cell, root, root tip, pistil, anther,floret, seed, pericarp, spike, stem, and callus.
 5. A wheat plantregenerated from the tissue culture of claim
 3. 6. A compositioncomprising a seed or plant part of wheat cultivar WB-112, and acultivation medium, wherein a representative sample of seed of wheatcultivar WB-112 has been deposited under ATCC Accession No. PTA-11955.7. The seed or plant part of claim 6, wherein the cultivation medium issoil or a synthetic medium.
 8. A wheat seed produced by crossing twowheat plants and harvesting the resultant wheat seed, wherein at leastone of the two wheat plants is the wheat plant of claim
 2. 9. A wheatplant, or a part thereof, produced by growing said seed of claim
 8. 10.An herbicide tolerant wheat plant produced by introducing a geneconferring herbicide tolerance into the plant of claim 2, wherein thegene confers tolerance to a herbicide selected from the group consistingof glyphosate, sulfonylurea, imidazolinone, dicamba, glufosinate,phenoxy proprionic acid, L-phosphinothricin, cyclohexone,cyclohexanedione, triazine, and benzonitrile.
 11. A pest or insectresistant wheat plant produced by introducing a gene conferring pest orinsect resistance into the wheat plant of claim
 2. 12. The wheat plantof claim 11, wherein the gene encodes a Bacillus thuringiensis (Bt)endotoxin.
 13. A disease resistant wheat plant produced by introducing agene conferring disease resistance into the wheat plant of claim
 2. 14.A wheat plant having modified fatty acid metabolism or modifiedcarbohydrate metabolism produced by introducing a gene encoding aprotein selected from the group consisting of glutenins, gliadins,phytase, fructosyltransferase, levansucrase, α-amylase, invertase andstarch branching enzyme or encoding an antisense of stearyl-ACPdesaturase into the wheat plant of claim
 2. 15. A method of introducinga desired trait into wheat cultivar WB-112, wherein the methodcomprises: (a) crossing a WB-112 plant, wherein a representative sampleof seed is deposited under ATCC Accession No. PTA-11955, with a plant ofanother wheat cultivar that comprises a desired trait to produce progenyplants wherein the desired trait is selected from the group consistingof male sterility, herbicide tolerance, herbicide resistance, insectresistance, modified fatty acid metabolism, modified carbohydratemetabolism, modified protein metabolism, modified phytic acidmetabolism, modified waxy starch content, modified protein content,increased tolerance to water stress and resistance to bacterial disease,fungal disease or viral disease; (b) selecting one or more progenyplants that have the desired trait to produce selected progeny plants;(c) crossing the selected progeny plants with the WB-112 plant toproduce backcross progeny plants; (d) selecting for backcross progenyplants that have the desired trait and essentially all of thephysiological and morphological characteristics of wheat cultivar WB-112listed in Table 1; and (e) repeating the crossing the selected progenystep and selecting for backcross progeny step two or more times insuccession to produce selected third or higher backcross progeny plantsthat comprise the desired trait and essentially all of the physiologicaland morphological characteristics of wheat cultivar WB-112 listed inTable
 1. 16. A wheat plant produced by the method of claim 15, whereinthe plant has the desired trait.
 17. The wheat plant of claim 16,wherein the desired trait is herbicide tolerance and the tolerance isconferred to an herbicide selected from the group consisting ofimidazolinone, dicamba, cyclohexanedione, sulfonylurea, glyphosate,glufosinate, phenoxy proprionic acid, L-phosphinothricin, triazine andbenzonitrile.
 18. The wheat plant of claim 16, wherein the desired traitis insect resistance and the insect resistance is conferred by a geneencoding a Bacillus thuringiensis endotoxin.
 19. The wheat plant ofclaim 16, wherein the desired trait is modified fatty acid metabolism,modified carbohydrate metabolism or modified protein metabolism, andsaid desired trait is conferred by a gene encoding a protein selectedfrom the group consisting of glutenins, gliadins, phytase,fructosyltransferase, levansucrase, α-amylase, invertase and starchbranching enzyme or encoding an antisense of stearyl-ACP desaturase. 20.The wheat plant of claim 16, wherein the desired trait is male sterilityand the trait is conferred by a gene that confers male sterility.
 21. Aproduct comprising the wheat plant or part thereof of claim 2, whereinthe product is selected from the group consisting of grain, flour,bread, cookies, cakes, crackers, noodles, pastries, baked goods,cereals, pasta, beverages, livestock feed, straw, constructionmaterials, and laundry starches.